Fig4B_RACE_STOML2_R2_AI33.sgd

HEK293T ishXrn1 cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with PA-X from the influenza A/Puerto Rico/8/1984 (H1N1) or vector, and luciferase reporters with inserted sequence from the STOML2 or YKT6 gene. Insertion size ranges from 15 bp to 99 bp. RNA was collected 24 hrs later and analyze by 5' RACE using a primer that binds to luciferase sequences.

HEK293T ishXrn1细胞经强力霉素（doxycycline）处理3~4天以诱导Xrn1基因敲低，随后分别转染甲型流感病毒A/Puerto Rico/8/1984 (H1N1)来源的PA-X蛋白或空载体，以及携带源自STOML2或YKT6基因的插入序列的荧光素酶报告基因载体。插入片段的长度范围为15 bp至99 bp。24小时后收集细胞总RNA，采用结合荧光素酶序列的引物通过5'末端快速扩增（5' RACE）技术进行分析。